IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS)

e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 4 Ver. III (Jul - Aug. 2015), PP 37-45
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Comparative Study of Using Sodium Chlorite and Tartaric Acid

Pretreatment on Waste Rice Straw for Enhanced Production of
Bioethanol
Arpan Kumar Basak1, Bunushree Behera1, Anuradha Mukherjee3,
Kaustav Aikat1, Amit Ganguly2, and Pradip Kumar Chatterjee2
1

Department of Biotechnology, National Institute of Technology Durgapur, Durgapur 713209, India

Thermal Engineering, CSIR Central Mechanical Engineering Research Institute, Durgapur 713209, India
3
NSHM, Durgapur Knowledge Campus, Arrah, Durgapur , 713212 ,India

Abstract: This is comparative study of developing bioethanol from age old waste rice straw (WRS) collected
from thatched roofs pretreated with tartaric acid and sodium chlorite separately. WRS of different ages,
collected from nearby village, they were dried, grinded and screened. Proximate analysis was conducted by
(Neutral detergent fibre) NDF and (Acid detergent fibre) ADF methods for estimating the hemicellulose,
cellulose and lignin content of variable age (not more than 6, 12, 24, 36 and 48 months), with least content
11.35% in 48 months age old sample. Pre-treatment was conducted on least lignin content biomass sample by
using different concentrations of tartaric acid and sodium chlorite under variable physical parameters such as
soaking temperature, soaking time and agitation speed which were optimized by response surface methodology
design tool, followed by moist heat treatment. The sample resulting best lignin degradation estimated by total
phenol content was subjected to saccharification by using both xylanase and cellulase. The time yielding
maximum hexose and pentose was 36th hour resulting in 494.8 mg/g and 274.9 mg/g glucose and 134.8 mg/g
and 124 mg/g xylose for tartaric acid and sodium chlorite pretreated samples respectively. After
saccharification the hydrolysate was collected and set for fermentation by Pichia stipitis. Ethanol yield was
obtained to be 20.226% and 25.467%.
Keywords: Response surface methodology, Sodium chlorite, Tartaric acid, Thatched, Waste rice straw (WRS)

I.

Introduction

Rice straw, commonly known as paddy straw is available widespread in and around India. It is the
vegetative part of Rice plant (Oryza sativa) which is cut during harvesting. The huge amounts of these straws
are burnt and are left in the fields till the next ploughing as Rice straw has its important role as soil improver.
Fresh rice straw is also used as a feed for livestock .It is also used as the interim material for idol structuring and
thatch making. So after few years these rice straws would add on to the bulk of waste and have absolutely no
role otherwise. They are either being thrown here and there or are being unwisely burnt off increasing the load
on greenhouse gases. [1,2]
So, an alternative reuse of rice straw from thatches are to be introduced which can be of utility to
mankind. Rice straw is an attractive lignocellulosic biomass for the production of bioethanol since it can be one
of the most abundant renewable resources available. It has several characteristics, such as high cellulose and
hemicelluloses content that can be readily hydrolyzed into fermentable sugars.
The choice of pretreatment methods plays an important role to increase the efficiency of enzymatic
saccharification thereby making the whole process economically viable Nature and its resources has resulted in
the scientific attraction in using biological renewable resources as a replacement to the use of non-renewable
resources which are rapidly depleting such as fossil fuels. Lignocellulosic biomass can be used as an alternative
to the automobile fuels due to its availability, abundance and relatively low cost. Lignocellulosic biomass from
old thatched roofs are considered to be effective as lignin content is very less compared to that of fresh rice
straw, on addition that fresh rice straw adds to the livestock and plays an important role in the food chain, but
waste rice straw on the other hand can be utilized for fermentation process for production of bioethanol as
renewable resource that stores energy from sunlight in its chemical bonds. It has been reported that ethanol
produced from lignocellulosic biomass resources has the potential to cut greenhouse gas emissions by 86% as it
is burnt more completely as compared to petrol. Ethanol has a much higher latent heat of vaporization (855
MJ/kg) than petrol (293 MJ/kg) as well as a higher octane number (99) than petrol (80100) as a result, preignition does not occur when ethanol is used. Moreover, it has no participation in the global warming crisis
because the carbon dioxide produced in the combustion process of ethanol is utilized by the plants for their
growth adding up to the carbon cycle balance in the nature.[2,3,4]
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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
Successive yield of cellulose and hemicellulose abundance was observed on physiochemical pretreatment of rice straw. Both acid hydrolysis and alkaline hydrolysis were conducted on rice straw for
progressive lignin degradation. On acid hydrolysis, lignin degradation was a success so as with alkali
pretreatment, but not a cost effective method for degrading lignin. Hence, the concept of using tartaric acid
fulfills the purpose of cost effectiveness. Tartaric acid was used on pre-treating wheat straw for lignin
degradation. Tartaric acid is found in remnants of tamarind, as we know abundance of both rice straw and
tamarind is drastic in southern parts of India, rural people can easily harvest tamarind and use it effectively for
acid hydrolysis of rice straw, which is definitely cost effective, and resulting in effective lignin degradation.
The present study deals with a modelling based statistical approach named Response surface
methodology (RSM) which was successfully implemented to find out the optimum conditions for
analyzing the amount of lignin yield by building up an experimental design. The fitting of the
responses extracted from design of experiments (DOEs) to a polynomial function is effectively done by
RSM. RSM includes statistical modelling to determine the interaction effects of the factors on the
response and serve as an effective tool for evaluating the optimum condition [5].
For the purpose of producing ligninase, ligninolytic fungi will have a higher activity when incubation
condition using submerged fermentation technique or Submerged fermentation (SMF) compared to solid
fermentation or Solid State Fermentation (SSF).

II.

Materials and methodology

2.1 Biomass estimation

Rice straw samples from thatched roofs were collected from settlings over a long period of time (6
months, 1 year, 3 years, 4 years approximately) from nearby village from reliable sources. These samples were
sun dried for a week and further moisture content were removed in a tray dryer for period 4 hours for
consecutive days at 60 C. These dried samples were finely grinded to powder and stored in air tight containers.
These powdered samples were used for experimental drives. The Hemicellulose and lignin content of the dried
and powdered biomass was estimated by NDF (Neutral Detergent Fibre) and ADF (Acid Detergent Fiber)
method. After estimating the lignin and hemicellulose content, theoretical value of cellulose and minerals were
obtained. To verify the actual amount of cellulose content present in waste rice straw of a particular age,
anthrone test was conducted as confirmatory quantitative test.[7]
2.2 Pretreatment
Finely powdered dried rice straw biomass were mixed with 10 mL of variable concentrations of tartaric
acid and sodium chlorite incubated for different incubation time, different agitation speed and at variable
soaking temperatures, these variable parameters were designed using a tool Design Expert which is based on
RSM (Response Surface Methodology) algorithm. After incubation, these samples were subjected to moist heat
(autoclaved at 15 psi, 121C, for 5 min. The series of treated samples were then filtered using Whatmanns filter
paper No. 1. Hydrolysate or filtrate was collected and concentration of lignin was determined by estimating total
phenol content by Folin-Cialcateau method and concentrations of reducing sugars both hexose and pentose
sugars were analyzed by DNSA and Phloroglucinol assay. [9]
2.4 Experimental design
structured, pre-designed and reliable

method

for

obtaining relationship between

various

parameters affecting a process ( ) and response of the process (Y). Central composite rotatable design
(CCRD) is among the principal experimental design technique used to analyse the interaction between
the process parameters. Combined effects of various process parameters such as soaking temperature,
soaking time , treatment time and agitation speed on reducing sugars and xylose yield after pretreatment
with sodium hydroxide was estimated using Response surface Methodology (RSM). The analysis was
done by Design Expert 9.0.3 trial software for optimization by Response Surface Methodology. The
software was used to estimate the responses of the dependent variable, regression analysis, graphical
analysis of the data obtained and to find out optimization efficiency.
The range of the independent parameters for the alkaline pretreatment by sodium hydroxide are
mentioned in Table. 1.
Table 1: Experimental range and levels of independent process variables for sodium
hydroxide pretreatment.
Factors
A
B
C

Name
Soaking Temperature
Soaking Time
Treatment Time

DOI: 10.9790/3008-10433745

Unit
C
h
min

Low
10
0
-10

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High
90
16
30

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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
D

Agitation

rpm

70

190

The input variables are scaled to coded levels based on the following equation Eq. (1)

xi =

Xi - X cp
Xi

i=1,2,3....k

(1)

where xi is a dimensionless parameter of the independent process variable, Xi indicates the

real value of the independent variable, Xcp implies the real value of an independent variable at the
centre point and Xi represents the step change in the real value of the variable i upon an unit
change in the dimensionless value of the variable i.
A second order polynomial equation Eq. (2) is used to estimate the relationship between the
independent and the experimental responses:
k

k-1

i=1

i=1 j=i+1

Y=o + i Xi + ii Xi Xi + ijXi X j

(2)

Analysis of variance (ANOVA) was used to find the statistical significance of the ratio of
mean square because of regression analysis and mean square due to the presence of residual error or
any noise. Fisher F test and P test was used to evaluate the significance of process parameters for a
given response. 3-D response surface graphs and contour plots were used to study and further confirm
the effect of significantly chosen parameters.
2.3 Saccharification
The pretreated samples with maximum phenol yield were subjected to enzymatic saccharification using
a combination of enzyme extracts cellulase and xylanase from Trichoderma reesei and Trametes versicolor
respectively. Both these enzymes were obtained from Sigma Aldrich. 103 U/g of cellulase and 650 U/g of
xylanase was used with substrate loading of 4% in Mendels media. It was then incubated at 50 C , at
(125-130 ) rpm for a period of 48 h. Sampling was done after 6 h interval and tested for the presence
of reducing sugars and pentose sugars by using different analytical techniques to optimize the entire
process with time. The presence of reducing sugars and pentoses as well as phenol content using different
analytical techniques to optimize the time taken to reach saturation point. Estimation of reducing sugar present
in the hydrolysate of biomass performed using DNS method, and the estimation of xylose was conducted by
modified Tollens method. The saccharification process with the maximum reducing sugars and xylose yield
was selected for further bioprocess. [8,9]
2.4 Fermentation
The microbial species Pichia stipitis (NCIM 3500) was used for fermentation purpose as it has the
ability to utilize both xylose and glucose. These microbial samples were obtained by the courtesy of National
Collection of Industrial Microorganisms. The inoculated flask was incubated at 30 oC on a rotating shaker at 125
rpm inside a BOD incubator. After 20 h of incubation, broth was collected in centrifuge tubes and each
centrifuge tubes were centrifuged at 10000 rpm for 10 mins. After centrifugation, the cell pellet formed at the
bottom of the tube were aseptically washed and suspended in sterile distilled water under pre-sterilized LAF
hood or a biosafety cabinet. For fermentation media (Ammonium chloride; 0.5 g/l, Potassium dihydrogen
phosphate; 2.0 g/l , Magnesium sulphateheptahydrate;0.5 g/l, Yeast extract; 1.5 g/l, Calcium chloride
dihydrate;0.1 g/l, Ferric chloride dihydrate;0.1 g/l, Zinc sulphateheptahydrate 0.001 g/l) was prepared and
autoclaved at 121.1 oC, 1.5 psi for 20 min. The hydrolysate obtained after saccharification by filtering the
biomass with muslin cloth and then Whatmanns filter paper 1 and it was supplemented into the media before
sterilization. The medium was inoculated with 10% inoculum of Pischia stipitis at pH 5.0 after sterilization
under LAF or aseptic condition and incubated at 30 C and 150 rpm. Sampling was done at regular intervals of
time then centrifuged with 15 minutes and tested for the presence of ethanol, reducing sugars and xylose.
Ethanol estimation was done by potassium dichromate method with the assistance of spectrophotometer. The
Estimation of reducing sugar present in the hydrolysate of biomass performed using DNS method, and the
estimation of xylose was done by modified tollens method. [9,11]

III.

Results and discussion

3.1 Proximate analysis

On conducting the NDF and ADF method the hemicellulose content and lignin content was estimated
for varying sample ages collected from old thatched roofs. Fig. 1 shows the hemicellulose content, Lignin
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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
content and cellulose content. It was observed clearly that with increase in age of the samples, its lignin content
decreases thereby exposing hemicellulose and cellulose, increasing their availability. The possibility of naturally
degrading lignin is maybe due to physiochemical or biochemical process or by both. The sample with least
lignin content was chosen for further bioprocessing. The lignin content was found to be 10.82% on 48 months
age old rice straw sample which was least among all. [1]

Figure 1: Represents the variation in rice straw components with age of biomass.
3.2 Saccharification
The samples pretreated with tartaric acid and sodium chlorite were subjected to saccharification
process, where the cellulose and hemicellulose content were enzymatically de-polymerized into their
monosaccharides, glucose, pentose and other reducing sugars with the involvement of cellulase (EC 3.2.1.4) and
xylanase (EC 3.2.1.8), majorly in order to break down cellulose and hemicellulose polysaccharides into is
monomer glucose and xylose. Xylose and glucose yield was optimized with time in hours (Table no. 5) and
compared with each pre-treated samples.[12]

Figure 2: Comparative saccharification of hemicellulose by xylanase for sodium chlorite and tartaric
acid pre-treated sample.
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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
Table 2: Comparative yields of pentose sugars after saccharification.
Time
(H)
6
12
18
24
30
36
42
48

Sodium
Chlorite
Pretreated Sample
60.55
70.313
81.4
98.96
121.1
124
93.8
72.3

Tartaric
Acid
Pretreated Sample
63.8
69.01
78.8
89.84
107.5
134.8
93.1
70.3

Figure 3: Comparative saccharification of cellulose content of sodium chlorite and tartaric acid
pretreated sample by cellulose
Table 3: Comparative yield of hexose sugars after saccharification
TIME
(h)

Sodium
Chlorite
Pretreated Sample

Tartaric Acid Pretreated

Sample

6
12
18
24

102.7
115.37
128
215.4

335.3
347
371
408.3

30
36

240.6
274.9

437.1
494.8

42
48

146
67.6

368
345

By estimating the yield of total reducing sugars and pentose sugars on various sampling taken on
variable time period while incubation lead to an optimized condition (Table no. 4) resulting maximum yield
(Figure no. 2) which was at 36th hour resulting in 494.8 mg/g glucose and 134.8 mg/g xylose for tartaric acid
pretreated sample. [12]
3.3 Fermentation
Fermentation was conducted for 12 hours, and based on graphical observations a relation can be drawn
that on gradual decrease of sugar contents there is an increase in ethanol production. By analyzing the graphical
representation of yield% versus time in hours, it is evident that Pichia stipitis is utilizing both pentose and
hexose for fermentation, as there is a peak increase in ethanol yield with an abyssal drop of sugar content.
Fermentation was set for 12 hours, and the maximum yield of ethanol 20.226%, was obtained on 9th hour of
fermentation sampling with an abyss drop of sugar content 14.5% and 12.9% hexose and pentose respectively
(Table no. 2). It is evident that Pischia stipitis utilizes both pentose and hexose sugar for fermentation, as there is
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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
gradual decrease of pentose sugar and hexose sugar and simultaneous increment in ethanol yield (Figure no. 3)..
The results obtained are in accordance with the ethanol yield of 21.1 g/l as obtained from rice straw in
other studies.[13]

Figure 4: Comparative ethanol % obtained for sodium chlorite and tartaric acid pre-treated rice straw biomass.
3.4 ANOVA Experimental design
The Model F-value of 561.76 implies the model is significant. There is only a 0.01% chance that an Fvalue this large could occur due to noise. Values of "Prob> F" less than 0.0500 indicate model terms are
significant. In this case B, D, AB, AD, BC, BD, CD, A^2, B^2, C^2, D^2 are significant model terms. Values
greater than 0.1000 indicate the model terms are not significant. If there are many insignificant model terms (not
counting those required to support hierarchy), model reduction may improve the model. The "Lack of Fit Fvalue" of 51911502.43 implies the Lack of Fit is significant. There is only a 0.01% chance that a "Lack of Fit Fvalue" this large could occur due to noise. Significant lack of fit is considered to be bad condition, which should
be analyzed and made to fit. Standard deviation was obtained to be 6.63 with R-Squared value 0.9981 and a
mean of 356.89 adjusted R-Squared value was 0.9963, a C.V. of 1.86%,
Predicted R-Squared 0.9890,
Adequate Precision was obtained to be 79.549. The "Pred R-Squared" of 0.9890 is in reasonable agreement with
the "Adj R-Squared" of 0.9963; i.e. the difference is less than 0.2. "Adeq Precision" measures the signal to noise
ratio. A ratio greater than 4 is desirable. The ratio of 79.549 indicates an adequate signal. This model can be
used to navigate the design space.
Table 4: Experimental range and levels of independent process variables for sodium hydroxide
pretreatment.
ANOVA for Response Surface Quadratic model
Analysis of variance table [Partial sum of squares - Type III]
Sum of
Source
Squares
df
Model
3.453E+005 14
A-SOAKING TEMP
6.483E-003
1
B-SOAKING TIME
17602.83
1
C-AGITATION SPEED
113.22
1
D-CONC. OF SODIUM CHLORITE 7886.20
1
AB
38704.31
1
AC
44.67
1
AD
6717.83
1
BC
9589.27
1
BD
4568.02
1
CD
2807.15
1
A^2
3372.85
1
B^2
19205.23
1
C^2
92463.02
1
D^2
1.941E+005 1
Residual
658.56
15
Lack of Fit
658.56
10
Pure Error
6.343E-006
5
Core Total
3.460E+005 29

DOI: 10.9790/3008-10433745

Mean
Square
24663.68
6.483E-003
17602.83
113.22
7886.20
38704.31
44.67
6717.83
9589.27
4568.02
2807.15
3372.85
19205.23
92463.02
1.941E+005
43.90
0.76
1.269E-006

F
Value
561.76
1.477E-004
400.94
2.58
179.62
881.57
1.02
153.01
218.41
104.05
63.94
76.82
437.44
2106.02
4420.17

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p-value
Prob> F
<0.0001
0.9905
<0.0001
0.1291
<0.0001
<0.0001
0.3291
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001

Significant

not significant

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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
The Model F-value of 88299.72 implies the model is significant. There is only a 0.01% chance that an
F-value this large could occur due to noise. Values of "Prob> F" less than 0.0500 indicate model terms are
significant. In this case A, B, C, D, AB, AC, AD, BC, BD, CD, A^2, B^2, C^2, D^2 are significant model
terms. Values greater than 0.1000 indicate the model terms are not significant. If there are many insignificant
model terms (not counting those required to support hierarchy), model reduction may improve your model. The
"Pred R-Squared" of 0.9999 is in reasonable agreement with the "Adj R-Squared" of 1.0000; i.e. the difference
is less than 0.2. "Adeq Precision" measures the signal to noise ratio. A ratio greater than 4 is desirable. The ratio
of 888.435 indicates an adequate signal. This model can be used to navigate the design space. The equation in
terms of actual factors can be used to make predictions about the response for given levels of each factor. Here,
the levels should be specified in the original units for each factor. This equation should not be used to determine
the relative impact of each factor because the coefficients are scaled to accommodate the units of each factor and
the intercept is not at the center of the design space. The surface plots were analyzed to determine the interaction
between process parameters on the total phenolic content for the tartaric acid hydrolysis of pretreated waste rice
straw biomass. The Response Surface plots shown in Figure nos. 4 (a), (b) and (c) which indicates the mutual
interaction between the process parameters are prominent.
Table 5: The data set analyzed by ANOVA (Analysis of variance) for response 1 lignin of tartaric acid
pretreated sample.
ANOVA for Response Surface Quadratic model
Analysis of variance table [Partial sum of squares - Type III]
Sum of
Mean
Source
Squares
df Square
Model
4.113E+005
14 29378.86
A-SOAKING TEMP
26582.43
1
26582.43
B-SOAKING TIME
4408.43
1
4408.43
C-AGITATION SPEED
7350.44
1
7350.44
D-Conc OF Tartaric acid
18.17
1
18.17
AB
10139.11
1
10139.11
AC
1.305E+005
1
1.305E+005
AD
999.44
1
999.44
BC
7291.26
1
7291.26
BD
666.58
1
666.58
CD
550.75
1
550.75
A^2
30150.96
1
30150.96
B^2
52979.88
1
52979.88
C^2
1.837E+005
1
1.837E+005
D^2
16389.15
1
16389.15
Residual
4.99
15 0.33
Lack of Fit
4.99
10 0.50
Pure Error
0.000
5
0.000
Cor Total
4.113E+005
29

F
Value
88299.72
79894.89
13249.76
22092.14
54.62
30473.63
3.922E+005
3003.86
21914.25
2003.43
1655.32
90620.29
1.592E+005
5.522E+005
49258.45

p-value
Prob> F
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001

significant

not significant

The equation in terms of actual factors for Sodium chlorite and tartaric acid pre-treated samples
(Equation 3 and 4) can be used to make predictions about the response for given levels of each factor. Here, the
levels should be specified in the original units for each factor. This equation should not be used to determine the
relative impact of each factor because the coefficients are scaled to accommodate the units of each factor and the
intercept is not at the center of the design space. [5]
Lignin=+1105.56701+20.03806* SOAKING TEMP-99.42441* SOAKING TIME-9.39037*
AGITATION SPEED-4994.96573* CONC. OF SODIUM CHLORITE-4.91835* SOAKING TEMP *
SOAKING TIME-3.71312E-003* SOAKING TEMP * AGITATION SPEED-20.49060* SOAKING TEMP *
CONC. OF SODIUM CHLORITE+0.54403* SOAKING TIME * AGITATION SPEED+168.96791*
SOAKING TIME * CONC. OF SODIUM CHLORITE+2.94348* AGITATION SPEED * CONC. OF
SODIUM CHLORITE+0.11089* SOAKING TEMP^2+26.46112* SOAKING TIME^2+0.028672*
AGITATION SPEED^2+8411.43667* CONC. OF SODIUM CHLORITE^2
Eq 3
Lignin = -1573.10912 +16.98686 * SOAKING TEMP +123.51256 * SOAKING TIME +15.99809 *
AGITATION SPEED +9.06614 * Conc OF Tartaric acid +0.62933 * SOAKING TEMP * SOAKING TIME 0.10034 * SOAKING TEMP * AGITATION SPEED +0.019759 * SOAKING TEMP * Conc OF Tartaric acid 0.23719* SOAKING TIME * AGITATION SPEED -0.16136 * SOAKING TIME * Conc OF Tartaric acid 6.51893E-003 * AGITATION SPEED * Conc OF Tartaric acid -0.082887 * SOAKING TEMP^2 -10.98737 *
SOAKING TIME^2 -0.040416 * AGITATION SPEED^2 -0.061111 * Conc OF Tartaric acid^2
Eq 4

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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
Design-Expert Software
Factor Coding: Actual
lignin (mg/g)
Design points above predicted value
Design points below predicted value
587.5

Design-Expert Software
Factor Coding: Actual
lignin (mg/g)
Design points above predicted value
Design points below predicted value
587.5

213.107

lig n in (m g /g )

X1 = A: SOAKING TEMP
X2 = B: SOAKING TIME

500

Actual Factors
C: AGITATION SPEED = 105
D: CONC. OF SODIUM CHLORITE = 0.3

400
300
200

500
400
300
200
100

100

60

0.4
0.35
0.3
0.25
D: CONC. OF SODIUM CHLORITE (mg/ml)
0.2

Design-Expert Software
Factor Coding: Actual
lignin (mg/g)
Design points above predicted value
Design points below predicted value
587.5
213.107

90

100

110

130

140

150

55

5.5
50

B: SOAKING TIME (hr)

C: AGITATION SPEED (rpm)

45

4.5

A: SOAKING TEMP (oC)

4 40

(b)

600

X1 = A: SOAKING TEMP
X2 = B: SOAKING TIME
Actual Factors
C: AGITATION SPEED = 105
D: CONC. OF SODIUM CHLORITE = 0.3

60

80

70

120

(a)

lig n in (m g /g )

Actual Factors
A: SOAKING TEMP = 50
B: SOAKING TIME = 5

600

600

lig n in (m g /g )

213.107
X1 = C: AGITATION SPEED
X2 = D: CONC. OF SODIUM CHLORITE

500
400
300
200
100

60

6
55

5.5
50

B: SOAKING TIME (hr)

45

4.5
4

A: SOAKING TEMP (oC)

40

(c)
Figure 5: RSM plots sodium chlorite: projecting the co-relation between variable parameters and lignin. (a)
Conc. of sodium chlorite with agitation speed; (b) Soaking time with soaking temperature; and (c) Soaking time
with soaking temperature.

Figure 6: RSM plots tartaric acid: projecting the co-relation between variable parameters and lignin. (a) Conc.
of tartaric acid with soaking time; (b) Agitation speed with soaking temperature; and (c) Soaking time with
soaking temperature.

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Comparative study of using sodium chlorite and tartaric acid pretreatment on waste rice straw for
IV.

Conclusion

It can be concluded that on an age old waste rice straw there was less lignin barrier compared to fresh
rice straw. The tartaric acid pretreatment resulted a better yield of ethanol compared to that of sodium chlorite
pretreatment. But on saccharification tartaric acid pre-treated sample resulted a better yield of xylose and
glucose compared to that of sodium chlorite pre-treated sample. Sodium chlorite can be considered as a better
solution than for delignification than tartaric acid. Lignin was removed effectively, as observed by estimating
the total phenol content. Optimization of parameters and yields for lignin were designed by Box-Behnken
design, this method also analyzed the relations between the parameters and yields. The experiments thus
conducted projects a clear image that for effective delignification sodium chlorite pre-treatment can be implied
resulting in a better ethanol yield using Pichiastipitis.

Acknowledgment

The authors are grateful to Director, CSIR Central Mechanical Engineering Research Institute,
Durgapur for constant support , encouragement and permission to publish the paper.

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